KR20210057068A - Biological sensing apparatus - Google Patents

Abstract

The present invention relates to a biological sensor 12 for detecting particles in a fluid. The living body detector 12 is included in an integrated circuit formed in a semiconductor manufacturing process and includes a particle detector 12 for sensing electrical properties and a flow device 30 for flowing a fluid. A particle detector is placed against the flow machine such that the detector detects the electrical properties of the particles in the fluid.

Description

Biometric sensing device {BIOLOGICAL SENSING APPARATUS}

The present invention relates to a sensing device and a method for sensing particles such as biological cells in a fluid, and to an analysis device including such a sensing device.

The analysis of the physicochemical properties of biological particles is a disease diagnosis. It is used for research and development, drug development, etc. There are known devices that do this analysis automatically and analyze a large number of particles per second at the same time. Typically, biological particles are suspended in a fluid before entering the analysis device, excite these suspended particles and detect their reaction. In a known analysis method, a fluorescent dye is added to the particle suspension, and then light is irradiated, and the light scattered by the particle suspension and the reflected light are detected and analyzed.

The present inventors have drawn attention to the disadvantages of the conventional analysis method of biological particles, specifically the large size of such an analysis device.

An object of the present invention is to make a living body sensing device for sensing particles in a fluid so portable that it can be carried by hand.

Another object of the present invention is to provide a biometric method for detecting particles in a fluid with a portable device.

The present invention in the biosensing device for detecting particles in a fluid:

Particle detectors that are included in the integrated circuit formed in the semiconductor manufacturing process to detect electrical characteristics; And

Including; a flow device for flowing a fluid,

There is provided a biosensing device in which the particle detector is disposed with respect to the flow device such that the particle detector detects electrical properties of particles in the fluid when the fluid passes through the flow device.

During use, a fluid sample, such as a biological sample, enters the flow unit and the fluid flows through the flow unit. The flow machine may, for example, form an open channel through which the fluid flows and use a pump. In addition, the flow device may have a structure in which the fluid flows by itself, for example, a structure in which fluid flows through a capillary phenomenon. Particle detectors are included in integrated circuits such as CMOS formed by semiconductor manufacturing processes. The particle detector is arranged to sense the electrical properties and to detect the electrical properties of the particles in the fluid passing through the flow machine. These particles are biological particles, such as cells, other structures including viruses, multicellular tissues, and bacteria. The detection of particles is analyzed by a method such as differentiating the cell samples from each other.

The particle detector can detect an amount corresponding to the dielectric constant of the particle. The particle detector can detect an amount corresponding to at least one of the actual and virtual dielectric constants of the particle. As will be described later, the particle detector can detect an electric field present in a fluid. Particle detectors can detect a response to a stimulus applied to a fluid. Particles, such as cells, react to and sense the stimulus applied in the form of a characteristic. Thus, particle detectors are used to detect the presence of particles in a fluid. Different particles respond to stimuli applied in different forms. Particle detectors can be used to characterize particles. For example, it is possible to differentiate the types of cells and determine characteristics such as the size or composition of the cells.

Stimulation in the form of an electric field to a fluid is suitable for sensing biological cells. In addition, it is possible to easily provide electrical sensors to an integrated circuit formed in a semiconductor manufacturing process, and in particular, a particle detector having a size corresponding to the size of a particle shape such as a biological cell can be provided.

Particle detectors can detect an electric field present in a fluid. Particles, such as cells, interfere with the electric field in response to the electric field. Therefore, the particle detector detects the presence of particles in the Yuchen according to the degree of disturbance of the electric field. Different particles have different forms of disturbing the electric field. Particle detectors can be used to determine the characteristics of particles. For example, the types of particles can be distinguished. The fluid may contain at least one electrically sensitive label. These labels are intended to improve response or enhance performance. The electric field generated by these labels can be changed or the response by the label can be used.

The particle detector includes a sensing array disposed relative to the flow device, so that the sensing array can detect an electric field present in the fluid during use. This sensing arrangement comprises at least a pair of electrodes, which electrodes are arranged with respect to each other and with respect to the fluid in the flow device to sense an electric field present in the fluid. These electrodes can be placed on one side of the fluid at right angles to the flow direction. The other electrode can be placed on one side of the fluid. These pairs of electrodes may be placed on either side of the fluid. In this case, the fluid detector may be included in a planar semiconductor integrated circuit such as a CMOS integrated circuit. The same size as the width or height of the electrode is less than 100㎛, 50㎛, 30㎛, 20㎛, 15㎛, 10㎛, 5㎛, 3㎛ or 1㎛, 0.5㎛, 1㎛, 5㎛, 10㎛ , 15㎛, 20㎛, may be greater than 30㎛ or 50㎛.

The particle detector includes a plurality of sensing arrays such as an electrode pair spaced apart from each other, and each sensing array detects particles, and the flow and particle detectors form a flow path through which particles flow, and when the particles flow along the flow path, the particles are continuous. It can be configured to be sensed by natural sensing arrangements.

The particle detector further includes a sensing circuit for receiving an input signal from the sensing array and providing an output signal, and the sensing circuit may detect electric charges present in the electrode. Specifically, the sensing circuit may include a capacitor that senses charge and converts the sensed charge into at least one of a voltage signal and a current signal. The sensing circuit has a high impedance input, providing the main sensing signal. Specifically, the sensing circuit may include an impedance buffer such as a field-effect transistor (FET). The FET provides a capacitive load on the electrode of the particle detector. The sensing circuit may provide a voltage signal or a current signal as an output signal. The sensing circuit may amplify the input signal. For example, the sensing circuit can convert and amplify the charge of the electrode into a voltage or convert it into a current and amplify it.

ESF (elerostatic field) can be used as a stimulus. Particle detector detects ESF. EDF (electrodynamic field) can also be used. For example, an electric field is applied in consideration of a bad electrochemical reaction to the electric field caused by biological cells and fluids, and if the electric field is too high, the cell may be destroyed. Because of this, the particle detector can detect EDF. EDF causes fewer bad electrochemical reactions to biological cells. For example, if the particle detector includes a sensing circuit, the sensing circuit may have sufficient bandwidth to respond to various signals sensed for the EDF.

The particle detector may include a stimulator that stimulates particles in the fluid. The stimulation of the particles causes a reaction in a form that is easy to detect by the above-described particle detector. The living body detector may be configured such that the stimulator and the particle detector operate at the same time. Specifically, the particle detector detects particles in the fluid, and at the same time, the stimulator also stimulates the particles in the fluid.

According to the present invention, the stimulator can apply an electric field to the fluid. When an electric field is applied to a fluid, a reaction is generated from the particle itself, and there is no need to use a label that accelerates the response to the stimulus. As described above, the stimulator to which an electric field is applied is easily constructed by a semiconductor manufacturing process such as CMOS. Power can be built into an integrated circuit equipped with a particle detector. In the semiconductor manufacturing process, power is provided in a size suitable for particles such as biological cells. If you have a label that is electrically sensitive to the fluid, you can properly configure the stimulator.

The power source is configured to apply an electric field to the fluid. Sometimes it is better to apply EDF to the fluid. Thus, the power source includes a voltage signal and a voltage source that generates a variable voltage signal. This voltage source applies a voltage signal to the electrodes, and the electrodes are placed against the flow machine such that an electric field is applied to the fluid inside the flow machine.

At least one of the sensing element and the stimulating element may configure the biosensor to be electrically isolated from the fluid in the flow device. This electrical isolation greatly reduces the charge flowing in the fluid. The reduction in charge maintains the integrity of the particles in the fluid and provides the accuracy of the measurement. Electrical isolation is achieved by placing an isolator between the stimulating element or the sensing element and the fluid. As an isolator, there is a dielectric layer included in an integrated circuit. Specifically, the dielectric layer includes a passivation layer such as silicon nitride. The isolator may also comprise a thermoplastic fluoropolymer such as polyvinylidene fluoride (PVDF). It has been found that particles such as cells in the fluid adhere less to the PVDF than to the passivation layer.

The stimulation device includes a magnetic pole array arranged with respect to the flow device, and this stimulation array applies an electric field to the fluid during use. The stimulation arrangement may include at least one electrode pair. The electrode pairs are arranged to apply an electric field to each other and to the fluid in the flow device. One of the electrode pairs may be disposed at right angles to the direction of the fluid and the other may be disposed on one side of the fluid. Both electrode pairs may be placed on one side of the fluid. In this case, it is suitable for planar semiconductor integrated circuits such as CMOS.

When the particle detector includes a sensing array including an electrode pair, at least one electrode among the electrode pairs of the sensing array may not be included in the electrode pair of the magnetic pole array. The biosensor can thus simultaneously stimulate and detect particles in the fluid. Specifically, the electrode pair of the stimulus array and the electrode pair of the sensing array may share the electrode. This method is suitable for single ended detection. Thus, the sensing arrangement may be a single-ended sensing arrangement. The electrode pair of the stimulus array and the electrode pair of the sensing array may not share an electrode, and this method is suitable for the differential sensing method. This sensing arrangement may be a differential sensing arrangement. Differential sensing arrangements are known to be less sensitive to changes in fluids, specifically media containing particles to be sensed. The differential sensing scheme reduces common mode signals. Reduction of the common mode signal is advantageous when the stimulus is a common mode signal and its response is single-ended. This method may be suitable for differential stimulation. This stimulus arrangement may be a differential stimulus arrangement. The differential stimulation arrangement is for applying a stimulation signal larger than the single-ended stimulation arrangement to the fluid.

The living body detector applies a predetermined stimulation signal to the fluid, and compares the stimulation signal with the response of the stimulation signal detected by the particle detector. This comparison involves the correlation of a given stimulus signal and response. The living body detector determines a time delay between the application of a predetermined stimulus signal and a reaction by the particles in the fluid, and determines a conversion function or applies a conversion function to the particles according to the time delay. The determined transformation function is used to characterize the particles.

The predetermined signal may include a pseudorandom noise signal. The living body detector may generate a pseudorandom noise signal. The pseudorandom noise signal may include an m-sequence. The m-sequence shows a flat power spectrum density over the desired operating bandwidth and can be easily provided through standard digital circuits, making it suitable for implementation in integrated circuits formed by semiconductor manufacturing processes such as CMOS.

The biological detector can detect the frequency component of the signal detected by the particle detector, and the frequency component is at least 1kHz, 10kHz, 50kHz, 100kHz, 250kHz, 500kHz, 1MHz, 5MHz, 10MHz, 25MHz, 50MHz, 75MHz, 100MHz, 250MHz, 500MHz, 750MHz, 1GHz, 1.25GHz, 1.75GHz, 2GHz, 2.5GHz, 2.75GHz or 5GHz. Usually, the frequency component is between 10 kHz and 100 MHz. When the frequency is 1 MHz or more, the particle detector can characterize the interior of at least one particle such as a biological cell. For frequencies above 1 kHz, it is necessary to characterize the particles with external characteristics such as the size of the biological cell. The power of the signal applied by the biosensor through the stimulator is at least 1 µm, 5 ㎼, 10 ㎼, 25 ㎼, 50 ㎼, 100 ㎼, 250 ㎼ or 500 ㎼. The stimulator included in the biosensor thus applies a stimulus signal such as an electric field having at least one corresponding frequency component. When the particle detector has a large number of stimulating elements and sensing elements, for example, a large electrode array addressed by high frequency, the addressing time for each stimulating element or sensing element may be shorter than the time required for stimulation or sensing. The biosensor is thus provided for the persistence of at least one of the stimulus signal and the particle detection. Specifically, the biosensor comprises a persistence circuit, such as a memory, and such a memory is configured to hold a state longer than a predetermined time and address each of a plurality of stimulus elements for a predetermined time. The persistence circuit is configured to apply a stimulus signal to the stimulus element for a time longer than a predetermined address time. When the persistence circuit includes a memory, examples of the memory include at least one of SRAM and DRAM. Each of the several outputs from the persistence circuit is electrically coupled to each of the plurality of stimulation elements.

When there are multiple electrode pairs of the biosensor, the electrode that stimulates the particle and the electrode that senses the response to the stimulation can be changed. The biosensor may be configured to apply a stimulus with a first electrode set and sense a reaction with a second electrode set, and then apply a stimulus with a second electrode set and detect a reaction with a third electrode set. This method is suitable when the electrodes are arranged in a row.

The particle detector of the biosensor may comprise an array of sensing elements, such as an electrode array. This arrangement extends in the direction of fluid flow by the flow device. Thus, it is possible to detect particles by means of continuous sensing elements for particles passing through the flow device. On the other hand, this arrangement may be perpendicular to the direction of the fluid. Specifically, it may have a two-dimensional arrangement by extending in the direction of the fluid and at a right angle to the direction of the fluid. The sensing element arrangement is for simultaneous detection and stimulation of multiple particles. Simultaneous detection of multiple particles involves performing multiple stimuli simultaneously. On the other hand, as will be described later, there may be an arrangement for the motion of the particles.

The biosensor may have an array of operating elements extending at right angles to the fluid direction, and the array of operating elements is arranged to exert a force on the particles inside the flow device. This force has a component perpendicular to the flow direction and moves the particles in a direction different from the flow direction.

The actuating element arrangement has electrodes, and these electrodes may form an arrangement. At least one electrode set in this arrangement is for at least one of motion, stimulation and sensing. For example, some electrodes may actuate at some time, stimulate at another time, and sense at another time. The biosensor may apply an operation signal to a plurality of operation elements arranged in a direction other than the fluid flow direction. The operating signal may be applied to a plurality of operating elements with at least one of different amplitudes and phases, respectively. When a motion signal having at least one of several amplitudes and phases is applied to other motion elements, a force is applied to the particle. These actions are used to manipulate or classify particles. Specifically, a group of particles or particles is moved to a desired array portion, specifically toward an array portion having stimulus and sensing electrodes having a size suitable for the size of the particle, and then sensed, and multiple portions of the array It can detect particles at the same time.

Since the biosensor can operate without a label, it can also act on unlabeled fluids such as fluorescent dyes or microbeads.

Biosensors may act on microbial samples, 500㎛, 250㎛, 200㎛, 150㎛, 100㎛, 50㎛, 25㎛, 10㎛, 5㎛, 2㎛, 1㎛, 500nm, 250nm, 100nm, 50nm , Particles having a size of less than 25 nm, 10 nm or 5 nm can be detected. On the other hand, the biosensor is 2nm, 5nm, 10nm, 25nm, 50nm, 100nm, 250nm, 0.5㎛, 1㎛, 2㎛, 5㎛, 10㎛, 25㎛, 50㎛, 100㎛, 150㎛, 200㎛ or 250 It is also possible to detect particles having a size larger than µm. The biosensor can be used for particles in a range that fits the size of at least one of the stimulus arrangement and the detection arrangement. Specifically, the size of at least one of the stimulus arrangement and the sensing arrangement corresponds to the size or size range of the particles. The sensing array or the stimulation array is made by a semiconductor manufacturing process and has a size capable of sensing or stimulating particles of a specific size or range.

The semiconductor manufacturing process may be a planar semiconductor manufacturing process, a metal oxide semiconductor process such as CMOS, a submicron semiconductor manufacturing process such as 0.35 μm CMOS, and a high voltage 0.35 micron CMOS manufacturing process.

The fluid is a liquid of Salon, and includes a liquid that carries particles, and specifically includes charge carriers such as salt molecules. The fluid may include phosphate buffered saline (PBS).

The fluidizer may form a main channel through which the fluid flows. The particle detector is arranged to detect particles in the main channel, and can be arranged on one or both sides of the fluid. Thus, elements such as the electrode of the particle detector can be placed in the gksWHr of the fluid. Meanwhile, the elements of the particle detector may be disposed on both sides of the fluid. The flow device includes a sample inlet for receiving a fluid sample by injection or the like, and this sample inlet is connected to the main channel. The flow machine has a sample outlet opposite the sample inlet, and the sample outlet is also connected to the main channel.

The sample outlet is for flowing fluid from the main channel. The flow machine has at least one other inlet disposed next to the sample inlet, specifically first and second different inlets disposed side by side respectively on both sides of the sample inlet. These other inlets communicate with the main channel. In use, a coating liquid such as PBS enters the other inlet and flows in the main channel, specifically, it flows in parallel with the fluid. The coating liquid is for registration of particles contained in the fluid using a fluid detection value, and helps to preserve the fluid passing through the flow device. At least one other outlet is arranged opposite the other inlet of the flow machine, and this other outlet is for flowing the coating liquid from the main channel.

The flow group is at least partially composed of a polymer such as polymethyl methacrylate (PMMA). The length of the flow machine is about 25mm and the width is about 10mm. Particle detector and flow device are formed separately. The particle detector and the flow device are arranged to be pulled together. The particle detector and the flow device are detachably connected to each other by fastening elements including a layer of silicon gaskets. Removable connections are made to facilitate exchange of fluids that have reached the end of their service life or require different types.

The biosensor may detect at least one of a photoelectric constant and a light transmittance. As will be described later, the particle detector may detect light emitted from a fluid. In addition to detecting the electrical properties of the fluid, the particle detector can also detect light emitted from the fluid. Particle detectors can also detect light emitted from a fluid. The emitted light is light that is transmitted, reflected, or scattered by the light applied to the fluid. On the other hand, the emitted light may be light re-emitted by a fluorescent dye in the fluid, and such fluorescent dye reacts to the re-emitted light according to the light penetrating into the fluid. The particle detector thus includes a light sensor positioned against the flow device, which detects the light emitted from the fluid. Specifically, the fluid detector may include a photosensitive electric circuit. Photosensitive electric circuits are stable in semiconductor manufacturing processes such as CMOS. The photosensitive electric circuit includes a photosensitive element such as a photodiode (PD). Since the photodiode occupies almost no area in the integrated circuit, it can be easily embedded in a high-density electrode array such as an electrode array described later.

The stimulus applied to the fluid may be electromagnetic radiation, light, or visible light. Visible light is suitable for particles such as biological cells. A stimulator can apply electromagnetic rays, such as light or visible light, to a fluid in addition to an electric field. Particles react to electromagnetic radiation. The fluid may include a compound such as a label that allows particles to react to electromagnetic rays, specifically, a fluorescent dye that re-emits light. Electromagnetic radiation sources include light sources such as LEDs or laser diodes. The electromagnetic ray source and its supporting device can be implemented in a semiconductor manufacturing process such as CMOS. The electromagnetic radiation source can be made separate from the integrated circuit comprising the particle detector.

The present invention also provides a bioanalytical device including such a biometric sensor. The bioanalytical device operates as a cell calculator, and further includes a control device. The control device controls the particle detector, for example, to control the detection arrangement when a particle is detected. If the biosensor is equipped with a stimulator, this control device determines whether the electrodes in the particle sensor and the stimulators are used for motion, stimulation or sensing. When the biosensor has an electrode arrangement, the control device determines the order of operation of the electrodes. The control unit consists of an electronic circuit such as a Field Programmable Gate Array (FPGA) or any suitable electronic device such as a microprocessor.

The bioanalytical device may include a flow inducing device, that is, a pump that guides a fluid passing through the flow device. The flow induction device is controlled according to the flow rate of the fluid passing through the flow device by the output of the particle detector. The control device receives the output of the particle detector and provides it to the flow induction device. The particle detector described above determines the flow rate of the flow device, which flow rate is determined by the control device. The particle detector includes a plurality of sensing arrays separated from each other, and each sensing array detects particles, and the flow rate is determined according to the separation between the sensing arrays and the detection time difference of the particles by each sensing array. Meanwhile, the characteristic value of the at least one particle described above may be compared with a predetermined threshold value, and the flow induction device may be adjusted according to the result. These characteristic values have reliability compared to a predetermined value. Specifically, when the reliability is lower than the threshold value, the flow induction device reduces the flow rate of the fluid to improve the characteristic value.

The bioanalytical device may include a processor. Since the processor receives the signal from the particle detector and converts it into a digital form, it has an AD converter and also includes a signal conditioning circuit such as an amplifier or filter. Processors include AD converters, amplifiers, filters, integrated circuits such as FPGAs, and dedicated integrated circuits such as Application Specific Integrated Circuits (ASICs).

The bioanalytical device may further include an analyzer. This analyzer makes decisions about the particles in the fluid by the output of the particle detector. For example, after performing AD conversion, it is determined according to the electric field measurement value by the particle detector. These decisions are made about the distinction of properties such as the density of the particles in the fluid and the size or composition of the particles. The analyzer consists of a PC, embedded microprocessor, and electronic devices such as FPGAs.

The present invention also

In the biosensing method for detecting particles in a fluid:

Allowing a fluid to flow through the flow device; And

It also provides a living body sensing method comprising; sensing the electrical properties of particles in a fluid passing through the flow machine using a particle detection device included in an integrated circuit formed by a semiconductor manufacturing process and disposed with respect to the flow machine.

1 is a block diagram of a bioanalyzing apparatus according to the present invention;
Fig. 2 is a block diagram of the biosensor of Fig. 1;
3 is a block diagram of an example of a bioanalyzing device;
Fig. 4 is a schematic diagram of a flow device of the biosensor of Fig. 2;
5 is a schematic diagram of a single-ended sensing-stimulating cell;
6 is a schematic diagram of a differential sensing-stimulation cell.

1 is a block diagram of a biometric apparatus 10 of the present invention, and includes a biometric sensor 12, a control processor 14, and an analyzer 16. The biosensor 12 receives an analyte in the form of a phosphate buffered saline (PBS 18) in which cells (which constitute a particle) are suspended. Instead of PBS, current detection methods that are independent of the suspended material can also be considered. As the analyte passes through the biosensor 12, it is activated, stimulated, and sensed (as detailed below) and then exits the outlet 20. The control processor 14 controls the operation of the electronic circuit inside the living body detector 12, for example, determines an operation sequence of the electronic circuit, applies a stimulus signal to an analyte, or processes signals sensed by the living body detector. Examples of processing include amplification of the sensed signal, AD conversion of the sensed signal, and storage of the converted signal. Although not shown, there may be a pump for passing the analyte through the biosensor 12.

For the stored detection signals, the analyzer 16 detects the presence of biological cells in the analyte, counts the number of cells, classifies cells, and determines cell characteristics such as cell dimensions and components, as well as analysis by the control processor 14. It also monitors changes in the control type of the sensor 12. The control processor 14 is composed of a separate AD converter, an amplifier, a memory, an electronic device including an integrated circuit such as an FPGA including a digital circuit and an ASIC including an analog circuit. The analyzer 16 consists of all suitable electronic devices including general purpose computers such as PCs, embedded microprocessors, and electronic circuits such as FPGAs. The control processor 14 and the analyzer 16 may be configured separately from each other or may be configured together in the same integrated circuit or a general-purpose computer.

Figure 3 is a prototype block diagram showing a CMOS ASIC, FPGA, PC, etc. that make up the elements of the bioanalyzer. A bioanalyzer suitable for manufacturing is made differently depending on the components within the same integrated circuit or module. For example, the functions of the bioanalyzer 16 are merged into the FPGA, and the functions of the control processor 14 are merged into the CMOS ASIC. A modification of the embodiment of FIG. 3 is also within the scope of the present invention.

2 is a block diagram of a biometric sensor 12 included in the bioanalyzing apparatus of FIG. 1. The flow device 30 of the biometric sensor 12 receives the analyte 18 and sends it to the outlet 20, which will be described in detail in FIG. 4. The living body sensor 12 has a two-dimensional electrode array 32. Although the flow device 30 and the electrode arrangement 32 of FIG. 2 are arranged side by side, the electrode arrangement 32 may be located above the flow device 30 (see FIG. 4). The control processing circuit 34 of the living body sensor 12 is electrically connected to the electrode array 32 and performs cell stimulation, sensing, and operation by the electrode array 320. The electrode array 32 and the processing circuit 34 ) Is made of a 0.35 micron CMOS process, etc.

Referring to the embodiment 40 of FIG. 3, both the electrode array 32 and the control processing circuit 34 are made of a CMOS ASIC 42. Each electrode array 32 has an electrode spacing of 2 μm, a size of 18 μm×18 μm, and a pitch of 20 μm. The thickness and dielectric constant of the ASIC's standard polyimide top layer are insufficient to properly couple the analyte and electrode array (32). According to the first method, the top layer of polyimide is removed by oxygen plasma surface treatment to reveal the silicon nitride layer underneath it, or the step of depositing the polyimide layer is omitted in the manufacturing process. Because of the hydrophilicity of the silicon nitride layer, the analyte surface is exposed to the maximum for the electrode. The second method is to remove the top layer of polyimide according to the previous method or to place a polyvinylidene fluoride (PVDF) layer on the exposed silicon nitride layer without depositing the polyimide layer. The PVDF layer is provided by spin coating the exposed silicon nitride layer. Meanwhile, a PVDF layer is formed as a separate layer overlying the ASIC to cover the exposed silicon nitride layer. After removing the top layer of polyimide or forming a PVDF layer, as described above, an ASIC 42 is deposited for the flow device 30 of FIG. 2 to analyze the electrode array 32 passing through the flow device 30 Let it meet with the substance. The control processing circuit 34 of the ASIC includes a binary to decimal decoder, a memory for matrix addressing of the electrode array 32, a global setting logic, and a bias circuit for a sensor output signal path. The global setting logic powers up all control lines by gating the control signals for the memory reset and global reset signals. The embodiment of FIG. 3 further includes a PCB supporting the connection of electric circuits supporting the ASIC 42. The circuit included in the PCB includes an FPGA 44 that provides various digital functions including generation of stimulation operation signals, addressing of each electrode in the electrode array 32, and communication with the bus (USB) module 46. vhg do. The FPGA 44 is Xilinz Spartan03, and functions such as generating a stimulus signal in the form of m-sequence through a linear feedback shift register. Meanwhile, an m-sequence may be provided as an external transmitter for easy change of m-sequence characteristics.

The PCB further includes a PLL (Phase Locked Loop) module 48 that provides integrated clock synthesis to the FPGA, and a switching power supply 50 for the FPGA. The PLL module 48 generates stable phase-locked clocks from a crystal oscillator to be used in the FPGA according to the operation of the clock management circuit included in the FPGA to induce an auxiliary clock signal from the main clock received from the PLL module. The switching power supply 50 includes switch mode power units that generate 3.3V and 1.8V from the 5V USB bus voltage received from the device. There is also an input signal control circuit 52 on the PCB, which is an FPGA or

It receives stimulation activation signals from an external transmitter, generates differential stimulation signals from single-ended stimulation signals, and provides a programmable gain amplification of the voltage swing of the stimulation operation signals. There is also an output signal control circuit 54 on the PCB, which executes various functions including programmable gain amplification/attenuation of initially amplified signals under the control of a PC, followed by gain-fixed low distortion amplification of sensed single-ended/differential signals. do. The output signal control circuit 54 has a 250 MSPS (M sample per second) AD converter, which receives the amplified and sensed signals, and converts the converted bitstream with a data output clock synchronous to the bitstream. To provide. The data output clock is sent to the FPGA 44 as a reference for the clock management circuit in the FPGA. The use of the data output clock as the reference value of the clock management circuit is to eliminate the delay in the converted bitstream and eliminate the time delay caused by the sensing circuit. Support circuits such as the power regulation circuit 56 are also on the PCB.

As described above, the USB module 46 of the PCB communicates with the Python/Numpy software for PC that executes the function of the analyzer 160 of Fig. 1. Specifically, the PC constitutes a circuit different from the ASIC 42 of the PCB. The PC receives the sensed data or data block obtained from the FPGA 44 and stored locally in real time, and decodes the received m-sequence cryptographic data by applying a high-speed Hadamard transform, etc. to quickly calculate the impulse response. The Hadamard transform is as follows:

Where Ψ'is the estimated output spectrum of the system under test, m is the sequence order, H is the Hadamard matrix, η is the measured m-sequence cipher response, and ξ ₁ and ξ ₂ are the correct m-sequence data to be used for the Hadamard matrix. These are the cipher/decryption matrices that convert in order ξ ₁ and ξ ₂ can be the same. The PC also provides frequency domain data by performing FFT (Fast Fourier Transform) on the decoded data. The frequency domain data is displayed for user translation. Frequency domain data provide characteristics such as the size or composition of biological cells in an analyte, thereby determining the properties of a particular cell type or distinguishing between different cell types. The PC counts the number of cells in the analyte, determines the cell density in the analyte according to the flow rate and volume of the flow device, and displays the cell number and density information to the user.

4 is a detailed view of the flow device 30 of the biosensor of FIG. 2, indicated by 70. The flow device 70 of FIG. 4 is formed of PMMA or silicon and has a length of 25 mm and a width of about 10 mm. The analyte flows through the main channel 72 of the flow device 70. The electrode array 74 is disposed on the main channel 72 and contacts the analyte flowing through the main channel. The CMOS ASIC is disposed on the electrode array 74, and the ASIC and the flow device 70 are detachably coupled to each other by a fastening element including a silicot gasket layer, so that the electrode and the main channel are properly disposed with respect to each other. I can. An analyte is injected into the sample inlet 76 of the flow machine by injection or the like, and the sample outlet 78 is on the opposite side. The sample inlet and outlets 76 and 78 are connected to the main channel 72. There are first and second different inlets 80 and 82 in this flower, each of which is located on either side of the sample inlet 76. The first and second other inlets 80 and 82 are also connected to the main channel 72, respectively. The flow machine also has first and second other outlets 84,86, these other outlets being arranged on opposite sides of the flow machine, respectively, and also communicate with the main channel. A coating liquid such as phosphate buffered saline (PBS) enters the first and second other inlets 80 and 82 during use, enters the sample inlet 76, and flows next to the analyte flowing in the main channel. The coating solution is for preserving the analyte flowing through the main channel while registering the cells contained in the analyte using the electrode array 74. The electrode array 74 selectively addresses internal electrodes to operate biological cells included in the analyte, and applies an operation signal to the selectively addressed electrodes. The electrode address and the application of the operation signal are performed by the electronic circuit described above. Specifically, a first sine wave signal or digital clock is applied to the electrodes of the first central row (row is in the same direction as the analyte) of the electrode array 74, and the second sinusoidal signal or clock is the first central row of the electrode array. It is applied to the electrodes of the second row on either side of. Since the sine wave signals or clocks applied to the electrodes in a row different from that of the first and second have at least one different phase and amplitude, a force acts on the biological cells inside the analyte, resulting in the flow direction of the analyte as shown in FIG. 4. Cells move at right angles. In FIG. 4, the cells exit the flow device 70 through the first other outlet 84. If the electrode pattern and the operation signal are different, it is possible to move a cell or cell group in a different desired direction by making a different type of operation, such as a different size of the electrode. Alternatively, it is possible to simultaneously detect a plurality of cells by different electrode array portions by moving other cells toward different portions of the electrode array before sensing. Each electrode in the electrode array can be selectively used for motion, stimulation, and sensing. Stimulation and sensing will be described later. The fluid flow in the fluidizers 30 and 70 may be reversed by stimulation, detection, and motion of particles contained in the fluid operating as described above.

Stimulation and sensing includes electric field stimulation and sensing. Biological cells interfere with the electric field in response to the electric field. It interferes with the electric field in different forms depending on the type and size of biological cells. Therefore, by detecting the disturbed electric field, it is possible to detect the presence of biological cells, determine the relative size of cells, and detect the type of cells. In some cases, analytes may contain electrically sensitive labels to enhance response or performance, such as latex microbeads coated with specific antibodies for use in basic electrical HIV testing; Microbeads coated with different antibodies of different materials, such as polystyrene or rubber, of different sizes and capable of identifying biological cell types; And conductive particles such as iron microbeads that absorb the electric field. Both electric field stimulation and sensing can be achieved within the CMOS ASIC of the type described above. Specifically, electrode arrays 32 and 74 are used for both electric field stimulation and sensing. Although each electrode may be used for stimulation and sensing at different times, electrode sets may be used for stimulation and sensing at the same time.

The pump of the bioanalytical device 10 of FIG. 1 is controlled according to the flow rate or characterization reliability of the analyte passing through the biometric sensor 12. In consideration of the flow rate of the analyte, separation between the electrode pairs of the biometric sensor 12 is performed, the control processing circuit 34 determines the movement time of the cells between the electrode pairs, and then the biometric sensor 12 is operated. After determining the speed of movement of the cells passing through, the pump is controlled according to this speed. For example, if the determined speed is lower than the threshold value, the control processing circuit 34 operates the pump to increase the flow rate. In consideration of the reliability of the characterization of the analyte, the control processing circuit 34 characterizes the biological cells and determines the reliability, then compares the determined reliability and the threshold value, and then controls the pump accordingly. If the determined reliability is lower than the threshold, the pump is adjusted to lower the flow rate to improve the characterization of biological cells.

5 is a sensing stimulation cell 100 for a single-ended operation, using electrodes 102 included in electrode arrays 32 and 74. The dielectric layer 104 disposed between the electrode 102 and the analyte 106 is for separating the electrode 102 from neighboring electrodes. Other electrodes 108 and 110 of FIG. 5 are for sensing and stimulating adjacent cells. The first and second multiplexers 112 and 113 of the multiplexer circuit used in the sensing stimulation cell 100 are for one of four states selected according to the first and second state selection bits 118 and 120. The output buffer 115 is driven by a select bit 125. The first memory bit 114 stores the state of the first state selection bit 118, and the second memory bit 116 stores the state of the second state select bit 120. The first and second memory bits 114 and 116 are for continuous application of the state selection bit to the first and second multiplexers 112 and 113, respectively. Since the address time of each electrode in the electrode array is shorter than the time required to apply a stimulus signal to this electrode and the time required to obtain a signal from this electrode, it is necessary to continuously apply the state selection bit. The address time of each electrode is short for most of the electrodes. The first and second memory bits 114 and 116 are configured in static random access memory (SRAM).

As described above, the first and second multiplexers 112 and 113 are for one of four states, and the address-sensing selection bit 125 includes the first and second state selection bits 118 and 120 as the first and second memory. It is for connecting to bits 114 and 116, respectively. For one of the four states, the electrode is addressed with the address-sensing select bit 125, and then the first and second state select bits 118 and 120 are stored as the first and second memory bits 114 and 116. . The configuration of the first and second memory bits 114 and 116 is for one of four states. In the first state, if both the first and second state selection bits 118 and 120 are 0, the electrode 102 is connected to a common ground potential through a switch. In the second state, if the first state selection bit 118 is 0 and the second state selection bit 120 is 1, the output buffer 115 operates, and the electrode 102 passes through the addressable output buffer 115 for detection. It is connected to the sensor output pin 122.

The sensor output pin 122 is electrically connected to a part of the control processing circuit 34 that processes the sensed signal. In the third state, when the first state selection bit 118 is 1 and the second state selection bit 120 is 0, the electrode 102 receives a stimulation input from the signal bus 124 for stimulation. The signal bus 124 is connected to a part of the control processing circuit 34 for generating a stimulus signal. In the fourth state, when the first state selection bit 118 is 1 and the second state selection bit 120 is 1, the electrode 102 receives an operation input from the signal bus 124. The signal bus 124 is connected to a portion of the control processing circuit 34 that generates an operation signal. That is, the signal bus 124 carries both a stimulus input and a motion input, and the second state selection bit 120 selects one of the two inputs according to the state. Each electrode in the electrode array has a multiplexer and a memory circuit shown in FIG. 5. Accordingly, each of the sensing stimulating cells 100 of FIG. 5 and all of the sensing stimulating cells in the ASIC can be used for one of motion, stimulation, and sensing. In consideration of the stimulation, a stimulation signal is further applied between one electrode and the adjacent electrode. In the embodiment of FIG. 5, a stimulation signal is applied between the second and third electrodes 108 and 110. When a stimulation signal is applied between the second and third electrodes 108 and 110, the first electrode 120 senses it. The pattern of stimulation and motion may vary from cycle to cycle, unlike above. The pattern of stimulation and motion is determined by the control processing circuit 34, which can be programmed to provide any number of desired patterns. For example, in the electrode array, electrode rows above and below an electrode row including electrodes addressed for sensing may be stimulated to stimulate electrodes to reduce direct application of a stimulation signal to the electrodes being sensed.

6 shows a configuration for the sensing stimulation cell 140, where adjacent first and second electrodes 142 and 144 are used, and as in the embodiment of FIG. 5, the first and second electrodes 142 and 144 and an analyte ( A dielectric layer 146 disposed between 148 is for separating these electrodes 142 and 144 from adjacent electrodes. The electrodes 150 and 152 are adjacent electrodes. Like the sensing stimulation cell 100 of FIG. 5, each of the electrodes 142 and 144 has a first multiplexer 154 and a second multiplexer 155, and the state selection of the first and second memory bits 156 and 157 associated with them The bits are for selection of one of the four states as described above.

In the differential amplifier 158 of the embodiment of FIG. 6, one input is connected to the first electrode 142 and the other input is connected to the second electrode 144. The two outputs of the differential amplifier 158 are connected to the first sensor output pin 164 and the second sensor output pin 166 through respective address-sensing output buffers 154, respectively. The first capacitor 160 is connected between inputs and outputs of the differential amplifier 158, and the second capacitor 162 is connected between inputs and outputs of the differential amplifier 158. The first switch 168 is connected to the first capacitor 160, and the second switch 170 is connected to the second capacitor 162. Both the first and second switches are controlled by the second memory bit 157 related to the first electrode 142, and when the sensing state is selected, the first and second switches are opened and the differential amplifier 158 is The differential signal sensed between the second electrodes 142 and 144 is transmitted to the first and second sensor output pins 164 and 166. Differential detection is achieved through two adjacent electrodes in this way. When the first memory bit 156 is 0 and the second memory bit 157 is 1, the first and second switches 168 and 170 are closed to short the first and second capacitors. Since the first and second capacitors become conductive due to a short circuit, the apparent voltage difference disappears.

Stimulation of the embodiment of FIG. 6 occurs as in the embodiment of FIG. 5 by the third and fourth electrodes 150 and 152 on both sides of the two adjacent electrodes 142 and 144 except for the following description. Except for the following description, as in the embodiment of FIG. 5, the first to fourth electrodes 142, 144, 150, and 152 are operated by selecting an appropriate state. The signal bus 172 of the circuit of Fig. 6 is limited to carrying only two signals. Accordingly, the circuit of FIG. 6 includes another (not shown) multiplexer for selecting a stimulus signal or an operation signal. The circuit of FIG. 6 may be configured to simultaneously apply stimulation and operation signals, or may be configured to perform stimulation in a single-ended flat state.

Methods other than electric field stimulation and sensing include optical stimulation and electro-optical sensing. Optical stimulation is achieved by radiating visible light from a light source such as an LED or laser diode to the flow device. In order to stimulate the light source, a suitable fluorescent dye is added to the analyte in a second form, and the absorbed and emitted light is also sensed in consideration of the fluorescent dye. This detection is made by photosensitive elements such as photodiodes contained within the ASIC. Taking this approach into account, it is also possible to combine electric field sensing and optical sensing. For example, the photosensitive devices may be dispersed between the electrodes, and the photosensitive devices may be selected by addressing and reading detection signals, as in the embodiments of FIGS. 5 to 6.

Claims (17)

In the bioanalytical device for detecting biological particles in a fluid:
A flow device having a channel through which a fluid flows;
A living body detector having a particle detector and a particle stimulator included in an integrated circuit formed in a semiconductor manufacturing process;
A flow inducer for flowing a fluid through the channel; And
Including; a controller for controlling the flow inducer so that the fluid containing the biological particles pass through the channel,
The integrated circuit is disposed in the channel, the particle stimulator includes an electric field source circuit and at least a pair of stimulation electrodes, the electric field source circuit is electrically connected to the stimulation electrodes, and the stimulation electrodes are disposed on the first side of the channel. , The particle detector includes a sensing circuit and at least a pair of sensing electrodes, the sensing circuit is electrically connected to the sensing electrodes, the sensing electrodes are disposed on the first side of the channel, and the electric field source circuit is through the stimulation electrodes. A first electric field is applied to at least one of the biological particles in the fluid, and the sensing circuit senses a second electric field of at least one biological particle generated by the application of the first electric field through the sensing electrodes, and the sensed second electric field Provide a corresponding output signal according to;
The controller controls the electric field source circuit to apply a first electric field to the biological particles in the fluid when the fluid flows through the channel, and when the fluid flows through the channel after the application of the first electric field by the electric field source circuit, the second Controlling the sensing circuit to sense the electric field and to provide a corresponding output signal according to the sensed second electric field;
A bioanalytical apparatus, characterized in that the first electric field applied to the fluid by the particle stimulator is an electrodynamic field (EDF). The bioanalytical device of claim 1, wherein the fluid comprises at least one electrically sensitive label. The bioanalyzing apparatus according to claim 1 or 2, wherein the at least one pair of stimulation electrodes and at least one pair of sensing electrodes are arranged side by side. The bioanalysis according to claim 3, wherein the stimulus electrodes and the sensing electrodes have sizes smaller than 100 μm, 50 μm, 30 μm, 20 μm, 15 μm, 10 μm, 5 μm, 3 μm or 1 μm. Device. The method of claim 3, wherein the particle detector includes a plurality of sensing electrode pairs spaced apart from each other, each sensing electrode pair senses a biological particle, and the channel forms a flow path through which the biological particles flow, and the particle detector is Bioanalyzing apparatus, characterized in that configured to detect the biological particles flowing along the flow path successive pair of sensing electrodes. The bioanalytical apparatus according to claim 5, wherein the particle detector comprises an array of sensing electrode pairs, and the sensing electrode pair array extends in a flow direction of a fluid passing through the channel and in a direction orthogonal to the flow direction. The bioanalyzing apparatus according to claim 1, wherein the particle detector senses a second electrodynamic electric field. The bioanalyzing apparatus according to claim 1, wherein at least one of the at least one pair of sensing electrodes and the at least one pair of stimulation electrodes is electrically isolated from the fluid in the channel. The bioanalyzing apparatus according to claim 1, wherein the particle detector detects differentially. The bioanalyzing apparatus according to claim 1, wherein the biosensor compares the first electric field and the second electric field. The method of claim 1, further comprising an array of operating elements extending in a direction perpendicular to the flow direction of the fluid passing through the channel, wherein the array of operating elements is disposed with respect to the channel so as to apply a force to the biological particles in the channel and from the biosensor. Bioanalyzing device, characterized in that it operates according to the corresponding signal of. The bioanalyzing apparatus according to claim 1, wherein the semiconductor manufacturing process is a metal oxide semiconductor manufacturing process. The bioanalyzing apparatus according to claim 1, further comprising an analysis arrangement receiving a corresponding output signal. The bioanalyzing apparatus according to claim 1, wherein the first electric field applied by the electric field source circuit includes a pseudorandom noise element. The bioanalyzing apparatus according to claim 1, wherein the at least one pair of stimulation electrodes and at least one pair of sensing electrodes have a common electrode. The method of claim 1, wherein the biosensor further comprises a sustain circuit for application of the first electric field, and the sustain circuit is addressed for a predetermined period of time and maintains the state for a longer period of time than the predetermined period of time to maintain the bioparticles in the fluid. Bioanalyzing apparatus, characterized in that the first electric field is continuously applied to at least one of the. In the biometric detection method for detecting bioparticles in a fluid:
Providing a biometric analysis device according to claim 1;
Allowing fluid to flow through the channel; And
A flow inducer is controlled to allow the fluid containing biological particles to flow through the channel, and the electric field source circuit is controlled to apply a first electric field to at least one of the biological particles in the fluid when the fluid flows through the channel, and the sensing circuit is controlled. And detecting a second electric field of at least one biological particle when the fluid flows through the channel and providing a corresponding output signal according to the sensed second electric field.

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